Structure of M1 RNA as determined by psoralen cross-linking
- 26 January 1988
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 27 (2) , 570-575
- https://doi.org/10.1021/bi00402a011
Abstract
The RNA moiety of ribonuclease P from Escherichia coli (M1 RNA) has been photoreacted with 4''-(hydromethyl)-4,5'',8-trimethylpsoralen (HMT) and long-wave UV light (320-380 nm) in a buffer containing 60 mM Mg2+, where the RNA moiety acts as a true catalyst of tRNA processing. Limited specific digestion and two-dimensional gel electrophoresis yield fragments cross-linked by HMT. By photoreversal of the isolated cross-linked fragments and enzymatic sequencing of the fragments, the positions of the cross-links have been elucidated. This method allows us to locate the cross-link to .+-. 15 nucleotides. Further assignments of the exact locations of the cross-links have been made on the basis of the known photoreactivity of the psoralen with different bases. Nine unique cross-links have been isolated in the M1 RNA including four long-range interactions. The short-range interactions are discussed here in detail.This publication has 13 references indexed in Scilit:
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